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Human RCC (human + rcc)
Selected AbstractsModulation of p21-activated kinase 1 alters the behavior of renal cell carcinomaINTERNATIONAL JOURNAL OF CANCER, Issue 9 2007Gerald C. O'Sullivan Abstract The p21-activated kinase 1 (Pak1) is a serine/threonine kinase whose activity is regulated by both Rho GTPases and AGC kinase family members. It plays a role in cytoskeletal remodeling and cell motility as well as cell proliferation, angiogenesis, tumorigenesis and metastasis. An involvement of Pak1 in renal cell carcinoma (RCC), which remains highly refractory to chemotherapy and radiotherapy, remains to be investigated. Pak1 expression, phosphorylation and kinase activity were examined in RCC cell lines and human tissue from normal and renal carcinoma. We report increased Pak1 expression and constitutive activity in the membrane and nucleus but not the cytoplasm of resected human RCC. To study a role for Pak1 in RCC, we developed 786-0 clones that expressed either a kinase-active Pak1L83,L86 2 different Pak1 dominant negative mutants, Pak1R299 and Pak1L83,L86,R299 or Pak1 siRNA. The expression of Pak1L83,L86 increased 786-0 proliferation, motility and anchorage independent growth, while the dominant negative mutants and Pak1 siRNA abrogated these effects. In addition, Pak1L83,L86 conferred resistance to 5-fluorouracil with a 40% ± 10% increase in cell viability. Conversely, Pak1L83,L86,R299, Pak1R299 and Pak1 siRNA conferred sensitivity with a 65.2% ± 5.5%, 69.2% ± 3.3% and 73.0% ± 8.4% loss in viability, respectively. Finally, Pak1 plays a role in renal tumor growth in vivo. Only 33% of mice developed tumors in the Pak1L83,L86,R299 group and no tumors developed from Pak1R299 cell challenge. Together these findings point to Pak1 as an exciting target for therapy of renal cancer, which remains highly refractory to existing treatments. © 2007 Wiley-Liss, Inc. [source] Tumor-infiltrating lymphocytes derived from human renal cell carcinoma: Clonal analysis of its characteristicsINTERNATIONAL JOURNAL OF UROLOGY, Issue 3 2008Tomoyuki Shimabukuro Aim: To assess the characteristics of activated tumor-infiltrating lymphocytes (TIL), we report the isolation, growth response, and functional analysis of a CD4 - CD8+ TIL-clone derived from human renal cell carcinoma (RCC). Methods: Bulk TILs were expanded from a human RCC and the lymphocytes were separated into a CD8+ enriched population. Subsequently, using the limiting dilution technique, a TIL clone was established and its growth response, phenotype and cytotoxic activity were analyzed. Results: A clone, T16-13, by day 94 numbering 1 × 107 cells, was harvested and characterized as a CD4 - CD8+ clone. On day 144, the cytotoxic activity of this clone against the autologous tumor was relatively high (2.3 ± 0.7 LU30/106 cells). Meanwhile, against allogeneic renal tumors, there was no cytotoxic activity (,0.1 LU30/106 cells). Conclusions: A TIL clone possessing modest autologous tumor-specific cytotoxicity can be isolated from human RCC. The characteristics analysis of various TIL clones may provide a better understanding of an RCC tumor microenvironment and may help to establish new modalities for the treatment of patients with metastatic kidney cancer. [source] 34 In vivo tumour hypoxia and carbonic anhydrase IX expression in xenografted human renal cell carcinoma animal models using probes, 124I-G250 pet, biodistribution and immunohistochemistry immunobiodistribution, and oxygen studiesBJU INTERNATIONAL, Issue 2006N. LAWRENTSCHUK Introduction:, Hypoxia stimulates angiogenesis and has been demonstrated in tumours where it correlates with resistance to treatment and poor prognosis. We have previously demonstrated hypoxia in human Renal Cell Carcinoma (RCC). The purpose of animal models was to further evaluate oxygen levels within RCC whilst also focusing on expression of the protein carbonic anhydrase IX (CA IX). This protein is stimulated by hypoxia and involved in angiogenesis and may be a potential tumour target for imaging and future therapies. Methods:, Balb/c nude mice had human RCC (SK-RC-52) xenografted subcutaneously. Tumours were grown to different volumes with oxygen levels measured. Further groups then had the radiolabelled monoclonal antibody 124I-G250 (that binds to CA IX) injected intravenously and had Positron Emission Tomography (PET), gamma counting and oxygen studies performed on days 0,1,2,3,5,7,10 and 14 post injection. Immunohistochemistry and autoradiography was also performed. Results:, An inverse relationship between tumour volume and hypoxia within the model was established (P < 0.001). Furthermore, CA IX was expressed by tumours with maximal uptake of 124I-G250 on days 2/3 by distribution with gamma counting that could be correlated with uptake on PET imaging. Conclusions:, The xenograft model confirms human RCC are hypoxic. Also, that the level of hypoxia is inversely proportional to tumour sise. A correlation was made between PET scanning with 124I-G250 and biodistribution within tumours by gamma counting confirming CAIX as an imaging and potential therapeutic target in RCC. [source] Establishment and characterization of seven human renal cell carcinoma cell linesBJU INTERNATIONAL, Issue 1 2000K.-H. Shin Objective,To establish human renal cell carcinoma (RCC) cell lines, and to investigate the cell phenotypes and molecular characteristics of human RCC cell lines and their corresponding tumour tissues. Materials and methods,Seven human RCC cell lines from pathologically proven RCCs were established. The histopathology of the primary tumours, in vitro growth characteristics and status of tumour suppressor genes, mismatch repair genes and microsatellite instability (MSI) were examined in cell lines and their corresponding tumour tissues. Five of the cell lines were derived from clear cells (SNU-228, -267, -328, -349, and -1272), one from granular cells (SNU-482), and one from mixed clear and granular cell types (SNU-333). The mutational status was compared for von Hippel-Lindau (VHL), p53, TGF-, type II receptor (TGF-,RII), hMSH2, and hMLH1 genes in the cell lines and their corresponding tumour tissues. The MSI status of the cell lines was determined by screening for adenine repeat sequences, e.g. BAT-25, BAT-26, and BAT-40. Results,All lines showed different doubling times and were confirmed by DNA fingerprinting analysis to be unique. Contamination by mycoplasma or bacteria was excluded. In two cell lines (SNU-349 and -1272) and their tumour tissues, mutations in the VHL gene were found. The SNU-267 line had a frameshift mutation in the p53 gene. A missense mutation of the TGF-,RII gene was detected in the SNU-1272 line and the corresponding tissue. Analysis of the repeat sequences showed one cell line (SNU-349) to have MSI and the other six to have microsatellite stability. As MSI is a hallmark of the inactivation of mismatch repair genes, the presence of hMSH2 and hMLH1 mutations was investigated in all seven cell lines. An inactivating homozygous single base-pair deletion of the hMLH1 gene was found only in the SNU-349 cell line and corresponding tissue. Moreover, a frameshift mutation within an 8-bp polyadenine repeat present in the hMSH3 coding region was found only in the MSI cell line and tumour tissue. Conclusion,These newly established RCC cell lines should provide a useful in vitro model for studies related to human RCC. The SNU-349 cell line should be especially useful for studies of MSI and mismatch repair-defective RCCs. [source] | ||||||||||